Pre-selection by double layer density gradient centrifugation improves the fertilising capacity of frozen-thawed, capacitated stallion sperm

Roser Morató, Juleide M.De Souza Soares, Guifré Orero, Teresa Mogas, Jordi Miró

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11 Citations (Scopus)

Abstract

The effect of combining double layer density gradient centrifugation (DL-DGC) with different capacitation treatments on the fertilising capacity of frozen-thawed stallion sperm was examined via a heterologous assay involving in vitro-matured, zona pellucida-free bovine oocytes. In a first experiment, aliquots of frozen-thawed stallion sperm were subjected to one of five capacitation treatments without DL-DGC - ionomycin at 1.0. μM, 0.1. μM, 0.05. μM or 0.01. μM, or caffeine at 200. μg/mL. The fertilising capacity of the semen was then assessed at 18. h by staining the above oocytes with 4,6-diamidino-2-phenylindole (DAPI) and examining for sperm penetration, the number of penetrated spermatozoa per oocyte, and male pronucleus formation. In a second experiment, aliquots of frozen-thawed stallion sperm were subjected to DL-DGC selection - or not - and then further subjected to the two best capacitation treatments (0.1. μM and 0.05. μM ionomycin). The fertilising capacity of the semen was then determined as above. The DL-DGC/capacitated sperm samples showed the highest mean penetration rates: 24.16% following capacitation with 0.1. μM ionomycin, and 12.21% following capacitation with 0.05. μM ionomycin. The capacitated but non-DL-DGC-selected sperm returned significantly lower values: 6.26% and 7.02% for the same ionomycin treatments respectively. These findings suggest that combining DL-DGC selection with ionomycin capacitation improves the fertilising capacity of frozen-thawed stallion sperm. © 2013 Elsevier B.V.
Original languageEnglish
Pages (from-to)62-68
JournalAnimal Reproduction Science
Volume139
Issue number1-4
DOIs
Publication statusPublished - 1 Jun 2013

Keywords

  • Capacitation
  • EquiPure TM
  • Frozen semen
  • Stallion

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